Real-time PCR quantitation of hepatitis B virus total DNA and covalently closed circular DNA in peripheral blood mononuclear cells from hepatitis B virus-infected patients
| Autor: | M.-C. Baclet, Sophie Alain, Véronique Loustaud-Ratti, A A Mazet-Wagner, François Denis |
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| Přispěvatelé: | Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Bactériologie, Virologie, Hygiène [CHU Limoges], CHU Limoges, Université de Limoges (UNILIM) |
| Rok vydání: | 2006 |
| Předmět: |
Hepatitis B virus
Hepatitis B virus DNA polymerase MESH: Reagent Kits Diagnostic medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity Virus 03 medical and health sciences 0302 clinical medicine Orthohepadnavirus [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases Virology medicine Humans 030304 developmental biology 0303 health sciences Hepatitis B Surface Antigens MESH: Humans MESH: Hepatitis B biology Reproducibility of Results MESH: Polymerase Chain Reaction cccDNA Hepatitis B medicine.disease biology.organism_classification [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology MESH: Sensitivity and Specificity MESH: DNA Viral MESH: Hepatitis B Surface Antigens 3. Good health MESH: Reproducibility of Results MESH: Hepatitis B virus MESH: Leukocytes Mononuclear Real-time polymerase chain reaction Hepadnaviridae DNA Viral [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology Leukocytes Mononuclear MESH: DNA Circular 030211 gastroenterology & hepatology Reagent Kits Diagnostic DNA Circular |
| Zdroj: | Journal of Virological Methods Journal of Virological Methods, Elsevier, 2006, 138 (1-2), pp.70-79. ⟨10.1016/j.jviromet.2006.07.019⟩ |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/j.jviromet.2006.07.019 |
| Popis: | International audience; It remains unclear whether hepatitis B virus (HBV) replicates in extrahepatic tissues, and particularly in peripheral blood mononuclear cells (PBMCs), which may serve as a reservoir for the maintenance of infection. A real-time PCR assay for the quantitation of total and covalently closed circular (ccc) HBV DNA in serum and in PBMCs was developed. This assay was highly sensitive (detection limit: 27 IU/mL), linear over a wide range (9 log10), and was displayed high inter- and intra-assay reproducibility for the quantitation of total DNA. Genotypes A to E were detected and the results were consistent with those obtained with the COBAS Amplicor HBV Monitor Test. The specificity of the methodology was increased by prior treatment with an enzyme that digests relaxed circular DNA, and the elimination of background signals from virus adsorbed to the surface of PBMCs. HBV DNA was detected in the serum and PBMCs of 12 HBsAg-positive patients, with less than 1% in the cccDNA form. In conclusion, the quantitation of total and ccc HBV DNA in PBMCs is potentially useful as a non-invasive marker, and may help to increase our knowledge of the natural history of hepatitis B. |
| Databáze: | OpenAIRE |
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